Abstract
The error-prone V(D)J recombination process generates considerable amounts of nonproductive immunoglobulin (Ig) pre-mRNAs. We recently demonstrated that aberrant Ig chains lacking variable (V) domains can be produced after nonsense-associated altered splicing (NAS) events. Remarkably, the expression of these truncated Ig polypeptides heightens endoplasmic reticulum stress and shortens plasma cell (PC) lifespan. Many questions remain regarding the molecular mechanisms underlying this new truncated Ig exclusion (TIE-) checkpoint and its restriction to the ultimate stage of B-cell differentiation. To address these issues, we evaluated the extent of NAS of Ig pre-mRNAs using an Ig heavy chain (IgH) knock-in model that allows for uncoupling of V exon skipping from TIE-induced apoptosis. We found high levels of V exon skipping in PCs compared with B cells, and this skipping was correlated with a biallelic boost in IgH transcription during PC differentiation. Chromatin analysis further revealed that the skipped V exon turned into a pseudo-intron. Finally, we showed that hypertranscription of Ig genes facilitated V exon skipping upon passive administration of splice-switching antisense oligonucleotides (ASOs). Thus, V exon skipping is coupled to transcription and increases as PC differentiation proceeds, likely explaining the late occurrence of the TIE-checkpoint and opening new avenues for ASO-mediated strategies in PC disorders.
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Acknowledgements
We thank the staff of our animal facility as well as C. Carrion for technical assistance with microscopy and cell cytometry. We also thank K. Rajewsky (The Max Delbrück Center for Molecular Medicine, Berlin, Germany) and S. Casola (Institute of Molecular Oncology Foundation, Milano, Italy) for providing DH-LMP2A mice. We are grateful to N. Diallon (MRC Clinical Sciences Centre, London, UK) for providing the pγSat plasmid and M. Alizadeh (UMR S 917, Rennes, France) for repertoire sequencing. This work was supported by grants from Fondation ARC (PJA 20161204724/PGA120150202338), INCa (PLBIO15-256), ANR (2017-CE15-0024-01), Ligue Contre le Cancer (comités Corrèze, Haute-Vienne), Fondation Française pour la Recherche contre le Myélome et les Gammapathies monoclonales (FFRMG) and Comité d’Organisation de la Recherche sur le Cancer du Limousin (CORC). M.O.A. and N.S. were funded by Société Française d’Hématologie (SFH) and Région Limousin, respectively.
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M.O.A. and N.S. designed and performed experiments, analyzed data and wrote the paper. J.-M.L. and A.M. contributed data to Fig. 6. O.M., S.L.N., and E.P. contributed 3D DNA-FISH data in Fig. 3. J.M. performed gene expression analysis in Fig. 4. M.V.A., J.S., C.S., and M.C. helped with the experiments and interpretation of the data. L.D. conceived the project, designed experiments, analyzed data and wrote the paper.
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Ashi, M.O., Srour, N., Lambert, JM. et al. Physiological and druggable skipping of immunoglobulin variable exons in plasma cells. Cell Mol Immunol 16, 810–819 (2019). https://doi.org/10.1038/s41423-018-0160-6
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DOI: https://doi.org/10.1038/s41423-018-0160-6
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